Stabilization of ozone-labile fluorescent dyes by thiourea

Abstract

The present invention provides compositions and methods for stabilization of fluorescent dyes. In particular, the present invention provides buffer systems comprising thiourea to protect against degradation of ozone-labile fluorescent dyes.

We claim: 
     
         1 . A composition comprising: i) a buffer; ii) a fluorescent dye; and iii) or more thioureas. 
     
     
         2 . The composition of  claim 1 , wherein one of said one or more thioureas comprises thiourea. 
     
     
         3 . The composition of  claim 1 , wherein said fluorescent dye comprises an ozone-labile fluorescent dye. 
     
     
         4 . The composition of  claim 3 , wherein said fluorescent dye comprises Cy5. 
     
     
         5 . The composition of  claim 1 , wherein said composition is formulated for use in molecular biology, biochemistry, biophysics, or cell biology applications. 
     
     
         6 . The composition of  claim 5 , wherein said composition is formulated for use in DNA microarray analysis. 
     
     
         7 . The composition of  claim 5 , wherein said composition is formulated for use in next generation nucleic acid sequencing. 
     
     
         8 . The composition of  claim 1 , wherein said composition comprises one or more of ammonium persulfate, formamide, boric acid, glycine, citric acid, HEPES (2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid), Triton (e.g., Triton X-100; polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether, octyl phenol ethoxylate, polyoxyethylene octyl phenyl ether, 4-octylphenol polyethoxylate,), SDS (sodium dodecyl sulfate), TWEEN (polyoxyethylene 20, 80, etc.), CHAPS (3[(3-Cholamidopropyl)dimethylammonio]-propanesulfonic acid), urea, MOPS (3-morpholinopropane-1-sulfonic acid), DTT (dithiothreitol; (2S,3S)-1,4-Bis-sulfanylbutane-2,3-diol), PIPES (1,4-Piperazinediethanesulfonic acid), EDTA, disodium salt, PBS Buffer (phosphate buffered saline), TEMED (N,N,N′,N′-Tetramethylethylenediamine), Tris HCl, sucrose, TBS Buffer (Tris, NaCl), TAE Buffer (Trizma, glacial acetic acid, EDTA), TBE Buffer (Tris, boric acid, EDTA), TG-SDS Buffer (Tris, glycine, SDS), phosphate buffer, magnesium chloride, magnesium sulfate, sodium chloride, sodium acetate, ammonium sulfate, and potassium chloride. 
     
     
         9 . A composition for use with fluorescent dyes comprising: i) a buffer and ii) one or more thioureas. 
     
     
         10 . The composition of  claim 9 , wherein one of said one or more thioureas comprises thiourea. 
     
     
         11 . The composition of  claim 9 , wherein said composition is formulated for use in molecular biology, biochemistry, biophysics, or cell biology applications. 
     
     
         12 . The composition of  claim 11 , wherein said composition is formulated for use in DNA microarray analysis. 
     
     
         13 . The composition of  claim 11 , wherein said composition is formulated for use in next generation nucleic acid sequencing. 
     
     
         14 . The composition of  claim 9 , wherein said composition comprises one or more of ammonium persulfate, formamide, boric acid, glycine, citric acid, HEPES (2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid), Triton (e.g., Triton X-100; polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether, octyl phenol ethoxylate, polyoxyethylene octyl phenyl ether, 4-octylphenol polyethoxylate,), SDS (sodium dodecyl sulfate), TWEEN (polyoxyethylene 20, 80, etc.), CHAPS (3[(3-Cholamidopropyl)dimethylammonio]-propanesulfonic acid), urea, MOPS (3-morpholinopropane-1-sulfonic acid), DTT (dithiothreitol; (2S3S)-1,4-Bis-sulfanylbutane-2,3-diol), PIPES (1,4-Piperazinediethanesulfonic acid), EDTA, disodium salt, PBS Buffer (phosphate buffered saline), TEMED (N,N,N′,N′-Tetramethylethylenediamine), Tris HCl, sucrose, TBS Buffer (Tris, NaCl), TAE Buffer (Trizma, glacial acetic acid, EDTA), TBE Buffer (Tris, boric acid, EDTA), TG-SDS Buffer (Tris, glycine, SDS), phosphate buffer, magnesium chloride, magnesium sulfate, sodium chloride, sodium acetate, ammonium sulfate, and potassium chloride. 
     
     
         15 . A method for stabilizing fluorescent dyes comprising placing said fluorescent dye in buffer comprising one or more thioureas. 
     
     
         16 . The method of  claim 15 , wherein said fluorescent dye comprises an ozone-labile fluorescent dye. 
     
     
         17 . The method of  claim 16 , wherein said fluorescent dye comprises Cy5. 
     
     
         18 . The method of  claim 15 , wherein one of said one or more thioureas comprises thiourea. 
     
     
         19 . A kit comprising one or more fluorescent dyes and buffer comprising one or more thioureas. 
     
     
         20 . The kit of  claim 19 , wherein at least one of said one or more fluorescent dyes comprises an ozone-labile fluorescent dye. 
     
     
         21 . The kit of  claim 20 , wherein one of said one or more fluorescent dyes comprises Cy5. 
     
     
         22 . The kit of  claim 19 , wherein one of said one or more thioureas comprises thiourea. 
     
     
         23 . The kit of  claim 20 , wherein said buffer is formulated for use in molecular biology, biochemistry, biophysics, or cell biology applications. 
     
     
         24 . The kit of  claim 23 , wherein said buffer is formulated for use in DNA microarray analysis. 
     
     
         25 . The kit of  claim 23 , wherein said buffer is formulated for use in next generation nucleic acid sequencing. 
     
     
         26 . The kit of  claim 19 , comprising one or more of ammonium persulfate, formamide, boric acid, glycine, citric acid, HEPES (2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid), Triton (e.g., Triton X-100; polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether, octyl phenol ethoxylate, polyoxyethylene octyl phenyl ether, 4-octylphenol polyethoxylate,), SDS (sodium dodecyl sulfate), TWEEN (polyoxyethylene 20, 80, etc.), CHAPS (3[(3-Cholamidopropyl)dimethylammonio]-propanesulfonic acid), urea, MOPS (3-morpholinopropane-1-sulfonic acid), DTT (dithiothreitol; (2S,3S)-1,4-Bis-sulfanylbutane-2,3-diol), PIPES (1,4-Piperazinediethanesulfonic acid), EDTA, disodium salt, PBS Buffer (phosphate buffered saline), TEMED (N,N,N′,N′-Tetramethylethylenediamine), Tris HCl, sucrose, TBS Buffer (Tris, NaCl), TAE Buffer (Trizma, glacial acetic acid, EDTA), TBE Buffer (Tris, boric acid, EDTA), TG-SDS Buffer (Tris, glycine, SDS), phosphate buffer, magnesium chloride, magnesium sulfate, sodium chloride, sodium acetate, ammonium sulfate, and potassium chloride.